FACS Fluorescence – Activated Cell Sorting

Applications include:

Immunophenotyping -4 color capability

Biomarker analysis

Cell viability (Annexin-PI)

Intracellular staining

Cell differentiation

Absolute cell counts

Calcium Flux

Cell cycle and ploidyROCK

Rho

FAK CrK

Pax PYK2

GTP Gα-O

GEFs

CDC42 Rac

MEKs

PAK MKKs

ASK1

JNK

P38 IP3R

PKC

BIMP1

BCL10

NFκB IκBs

P

IKKs

IκBsDegradation

Src

SOSGRB2SHC

RasGEFs

Raf1

MEKs

ERKs

PLC-β

DAG

NFκB

IKKs

PκB

IP3

Ca+2Ca+2

Ca+2

Calm

P

Caln

NFAT

NFAT

P

PI3K

Gβ Gγ

LARG

BRaf

MEKs

ERKs

cAMP

EPACPKA

PDZRhoGEF

Gα-1 GTP

Rap1

p115RhoGEF

RT

KS

CREB

Gα12/13

SRF

GTP

CREB

SRF ELK-1

c-Jun

c-Fos

STAT

NFκBNFAT

Gene Expression

Gα Gβ

© 2013 CDT all rights reserved

CON

LFU

ENCE

DIS

COVE

RY T

ECH

NO

LOG

IES

| TE

CHN

ICAL

BRI

EFTECHNICAL BRIEF:

FACSFluorescence - Activated

Cell SortingD I S C O V E R Y T E C H N O L O G I E S

FACS is an essential tool for in-depth cell analysis. With the capacity to simultaneously measure multiple parameters on hundreds of individual cells per second, FACS is a power-ful technology with a wide variety of applications in pharma-ceutical development. The Confluence Discovery Technologies (CDT) team has a wide range of experience in developing and validating FACS assays to suit many different requirements. Our highly trained and knowledgeable scientists will assist with experi-mental design to develop and validate assays to meet client-specified criteria. We also perform data analysis and interpretation of FACS data to enable our clients to obtain the best insights from large, complex data outputs. For assay design, development, validation, sample and data analysis, our experience in research and clinical devel-opment projects enables us to be an ideal FACS service part-ner.

Assay DevelopmentThe CDT team can advise on the most suitable assay proce-dure depending on the requirements of the client. Including:

•Sample type (e.g., whole blood, separated PBMC, serum, cultured cells)

•Stability of marker(s)/analyte(s) being measured

•Appropriate fluorochromes, dyes and conjugates to get the clearest data from your samples

•Incubation temperatures and periods most suited to the matrix and/or analytes

•Most suitable controls to monitor assay performance

•Appropriate QC checks on instrumentation to ensure cytometers are performing reproducibly

ROCK

Rho

FAK CrK

Pax PYK2

GTP Gα-O

GEFs

CDC42 Rac

MEKs

PAK MKKs

ASK1

JNK

P38 IP3R

PKC

BIMP1

BCL10

NFκB IκBs

P

IKKs

IκBsDegradation

Src

SOSGRB2SHC

RasGEFs

Raf1

MEKs

ERKs

PLC-β

DAG

NFκB

IKKs

PκB

IP3

Ca+2Ca+2

Ca+2

Calm

P

Caln

NFAT

NFAT

P

PI3K

Gβ Gγ

LARG

BRaf

MEKs

ERKs

cAMP

EPACPKA

PDZRhoGEF

Gα-1 GTP

Rap1

p115RhoGEF

RT

KS

CREB

Gα12/13

SRF

GTP

CREB

SRF ELK-1

c-Jun

c-Fos

STAT

NFκBNFAT

Gene Expression

Gα Gβ

© 2013 CDT all rights reserved

CDTv1.8.13

Confluence Discovery Technologies, Inc.4320 Forest Park Avenue

Saint Louis, MO 63108Phone: 314-932-4032 x305

Fax: 314-932-4038www.confluencediscovery.com

CDTinfo@confluencestl.com

D I S C O V E R Y T E C H N O L O G I E S

CAPABILITIESD I S C O V E R Y T E C H N O L O G I E S

CON

LFU

ENCE

DIS

COVE

RY T

ECH

NO

LOG

IES

| CA

PABI

LITI

ES

Leaders in biochemistry and mechanistic enzymology, our senior team has access to a suite of capabilities and technologies enabling: • Drug:Target interaction • Enzyme:Inhibitor mechanisms • Direct binding kinetics • Compound mechanism of action

Con�uence o�ers a spectrum of cell and molecular biology capabilities: • Identi�cation of biologically important signal transduction pathways • Quantitation of e�cacy and selectivity • Understanding safety issues linked to pathways • De-convolution to isolate pathway nodes • Evaluation of multiple targets within the pathway • Identi�cation of key pathway point(s) for therapeutic intervention • Target validation, cloning, expression, and puri�cation • Custom assay development • High-throughput screening

Our team delivers when “OFF the SHELF” is not an option: • Assay development, optimization, and validation • Multiplex technology platforms and instrumentation • Complex data analysis • Specialty screens for preclinical technical application • Trouble shooting and problem solving

Our team provides clients with the expertise to: • Identify target, mechanism, and disease biomarkers • Select the appropriate biomarkers for a drug discovery program • Execute biomarker assay design, development, validation, and interpretation for pre-clinical and clinical studies • Seamless assay transfer from pre-clinical to clinical studies

Our medicinal and computational chemists are experts in designing and delivering novel chemical entities through: • Schrödinger® enabled computational drug design • High throughput hit prioritization • Chemical series prioritization • Structure activity relationship generation • Custom synthesis